Single-cell RNA-sequencing reveals a distinct population of proglucagon-expressing cells specific to the mouse upper small intestine.
Single-cell RNA-sequencing reveals a distinct population of proglucagon-expressing cells specific to the mouse upper small intestine.
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DOI：
10.1016/j.molmet.2017.07.014
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发表时间：
2017-10
影响因子：
8.1
通讯作者：
Gribble FM
中科院分区：
文献类型：
--
作者：
Glass LL;Calero-Nieto FJ;Jawaid W;Larraufie P;Kay RG;Göttgens B;Reimann F;Gribble FM
鉴定产生胰高血糖素样肽-1的肠前胰高血糖素原表达（PPG）细胞亚群及其相关的感觉受体表达谱，从而能够发现靶向这些细胞群治疗糖尿病和肥胖症的治疗策略。我们对通过流式细胞术从3只GLU-Venus小鼠的小肠上部纯化的PPG细胞进行单细胞RNA测序。在低深度对来自2只小鼠的细胞进行测序，并且在高深度对来自第三只小鼠的细胞进行测序。来自234个PPG细胞的高质量测序数据用于通过tSNE分析鉴定簇。进行qPCR以比较簇特异性基因的纵向和隐窝/绒毛位置。免疫荧光和质谱法用于确认蛋白质表达。PPG细胞形成3个主要簇：具有经典L细胞的典型特征的一组，包括Gcg和Pyy的高表达（占所有PPG细胞的51%）;与表达Gip的K细胞重叠的细胞类型（14%）;以及表达Tph 1和Pzp的独特簇，其主要位于近端小肠绒毛中并共同产生5-HT（35%）。G蛋白偶联受体的表达在簇之间不同，表明细胞类型受到差异调节，并且具有差异靶向性。我们的研究结果支持了一个新的概念，即许多肠内分泌细胞群是高度重叠的，单个细胞产生一系列先前分配给不同细胞类型的肽。这些簇中不同的受体表达谱突出了增加肠道激素分泌以治疗糖尿病和肥胖症的潜在药物靶点。单细胞RNAseq将上小肠PPG细胞分成3个主要簇。PPG细胞簇表现出不同的激素和受体的表达谱。PPG细胞产生多种可通过质谱法检测的激素。一种新的小鼠PPG细胞簇表达标记物Tph 1和Pzp。
To identify sub-populations of intestinal preproglucagon-expressing (PPG) cells producing Glucagon-like Peptide-1, and their associated expression profiles of sensory receptors, thereby enabling the discovery of therapeutic strategies that target these cell populations for the treatment of diabetes and obesity. We performed single cell RNA sequencing of PPG-cells purified by flow cytometry from the upper small intestine of 3 GLU-Venus mice. Cells from 2 mice were sequenced at low depth, and from the third mouse at high depth. High quality sequencing data from 234 PPG-cells were used to identify clusters by tSNE analysis. qPCR was performed to compare the longitudinal and crypt/villus locations of cluster-specific genes. Immunofluorescence and mass spectrometry were used to confirm protein expression. PPG-cells formed 3 major clusters: a group with typical characteristics of classical L-cells, including high expression of Gcg and Pyy (comprising 51% of all PPG-cells); a cell type overlapping with Gip-expressing K-cells (14%); and a unique cluster expressing Tph1 and Pzp that was predominantly located in proximal small intestine villi and co-produced 5-HT (35%). Expression of G-protein coupled receptors differed between clusters, suggesting the cell types are differentially regulated and would be differentially targetable. Our findings support the emerging concept that many enteroendocrine cell populations are highly overlapping, with individual cells producing a range of peptides previously assigned to distinct cell types. Different receptor expression profiles across the clusters highlight potential drug targets to increase gut hormone secretion for the treatment of diabetes and obesity. Single cell RNAseq divides upper small intestinal PPG-cells into 3 major clusters. PPG-cell clusters exhibit distinct expression profiles for hormones and receptors. PPG-cells produce a variety of hormones detectable by mass spectrometry. A new mouse PPG-cell cluster expresses the markers Tph1 and Pzp.
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影响因子：
8.1
作者：
Lam BYH;Cimino I;Polex-Wolf J;Nicole Kohnke S;Rimmington D;Iyemere V;Heeley N;Cossetti C;Schulte R;Saraiva LR;Logan DW;Blouet C;O'Rahilly S;Coll AP;Yeo GSH
通讯作者：
Yeo GSH
影响因子：
24.5
作者：
Symonds EL;Peiris M;Page AJ;Chia B;Dogra H;Masding A;Galanakis V;Atiba M;Bulmer D;Young RL;Blackshaw LA
通讯作者：
Blackshaw LA
影响因子：
4.8
作者：
Habib, Abdella M.;Richards, Paul;Gribble, Fiona M.
通讯作者：
Gribble, Fiona M.
影响因子：
3
作者：
Hodge, Daryl;Glass, Leslie L.;Reimann, Frank
通讯作者：
Reimann, Frank
影响因子：
29
作者：
Reimann F;Habib AM;Tolhurst G;Parker HE;Rogers GJ;Gribble FM
通讯作者：
Gribble FM